EP0541625A1 - Systeme de formation d'images a foyer commun utilise en microscopie. - Google Patents

Systeme de formation d'images a foyer commun utilise en microscopie.

Info

Publication number
EP0541625A1
EP0541625A1 EP91913800A EP91913800A EP0541625A1 EP 0541625 A1 EP0541625 A1 EP 0541625A1 EP 91913800 A EP91913800 A EP 91913800A EP 91913800 A EP91913800 A EP 91913800A EP 0541625 A1 EP0541625 A1 EP 0541625A1
Authority
EP
European Patent Office
Prior art keywords
combination
slit
reflective
shaped
mask
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP91913800A
Other languages
German (de)
English (en)
Other versions
EP0541625B1 (fr
Inventor
John Graham White
William Bradshaw Amos
James Michael The Gate Fordham
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Medical Research Council
Original Assignee
Medical Research Council
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Medical Research Council filed Critical Medical Research Council
Publication of EP0541625A1 publication Critical patent/EP0541625A1/fr
Application granted granted Critical
Publication of EP0541625B1 publication Critical patent/EP0541625B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G02B21/0048—Scanning details, e.g. scanning stages scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B17/00—Systems with reflecting surfaces, with or without refracting elements
    • G02B17/02—Catoptric systems, e.g. image erecting and reversing system
    • G02B17/06—Catoptric systems, e.g. image erecting and reversing system using mirrors only, i.e. having only one curved mirror
    • G02B17/0647—Catoptric systems, e.g. image erecting and reversing system using mirrors only, i.e. having only one curved mirror using more than three curved mirrors
    • G02B17/0652—Catoptric systems, e.g. image erecting and reversing system using mirrors only, i.e. having only one curved mirror using more than three curved mirrors on-axis systems with at least one of the mirrors having a central aperture
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0028—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders specially adapted for specific applications, e.g. for endoscopes, ophthalmoscopes, attachments to conventional microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B26/00—Optical devices or arrangements for the control of light using movable or deformable optical elements
    • G02B26/08—Optical devices or arrangements for the control of light using movable or deformable optical elements for controlling the direction of light
    • G02B26/10—Scanning systems

Definitions

  • This invention relates to a confocal imaging system for microscopy and, in particular, to such a system which utilises slit scanning.
  • Confocal imaging systems have become established as an effective means of eliminating out-of-focus interference in optical microscopy.
  • the illumination in the object plane may be in the form of a single point, an array of points, a single line or an array of lines.
  • a mask with the same geometrical form as the illuminating pattern is incorporated in a plane conjugate with the object plane, so that only signals from the immediate vicinity of the illuminated regions are admitted through the mask into the viewing system. In this way, interfering signals emanating away from the illuminated regions are rejected.
  • a complete image is built up by scanning the illuminating pattern in such a way as to cover the whole of the area of the object plane, whilst keeping the mask in register with the illumination.
  • a confocal imaging system comprising means for forming a slit-shaped or bar-shaped illuminating beam, or an array of such beams, a beam- splitting means by which the light is directed into an optical microscope having an objective lens, and an optical scanning means means whereby the illuminating beam or beams is or are caused to scan and be directed into the objection lens of the microscope in order to scan the specimen, such optical scanning means consisting of a wholly reflective optical system for both focussing and scanning .
  • the same reflective optical system causes the emitted or reflected beam to be descanned (ie brought to a stationary state) and focussed upon a stationary confocal mask such as a slit of variable width, or an array of such slits.
  • the stationary slit image delimited by the mask can then be rescanned by a second wholly reflective optical system and brought to focus within an eyepiece or upon the photosensitive surface of a video camera as a two- dimensional image.
  • the first-mentioned reflective system and likewise the second reflective system, consists of a modified Offner autocolli ation system, in which the inner element of the Offner system is caused to oscillate. This modification converts the known Offner relay into a, unique reflective scanning system.
  • the confocal imaging system is exemplified in the following description, making reference to the single figure of drawings, in which a preferred system is shown schematically.
  • the system is interposed between the objective (OBJ) and the eyepiece (EYE) of a conventional microscope. It includes two Offner reflecting autocollimation units (A1,B1 and A2,B2) .
  • An Offner autocollimation system consists of a pair of mirrors with radii of curvature in the ratio 1:2.
  • the mirror of smaller radius is convex, the other concave.
  • this system functions as a 1:1 optical relay if light is reflected through it in the manner shown in the drawing.
  • the Offner system consisting of mirrors Al and Bl focusses the slit C at the plane shown as a dotted line in the eyepiece (EYE).
  • the other Offner system consisting of mirrors A2 and B2, creates conjugate foci in the plane OP and on the median plane MP at the slit C and within the lens LI.
  • the value of the Offner system in the present context is its ease of fabrication, total achromatism and freedom from primary aberrations.
  • Illumination is injected via a beam splitter (BS) .
  • the light could be provided by an illuminated slit.
  • a slit is not used. Instead a parallel laser beam is passed through a cylindrical lens (CL) and then through a conventional spherical lens ( 2) to provide a focussed line of laser light in the plane OP which is conjugate with the object plane.
  • the illuminating beam is directed through the Offner unit proximal to the microscope (B2, A2) .
  • the Offner units are modified in that the convex mirrors Bl and B2 are each caused to oscillate about an axis perpendicular to the plane of the diagram, whereby the beam is caused to scan.
  • Both of the convex mirrors shown in Figure 1 lie in the aperture planes in relation to the microscope rather than in image planes and they are therefore -ideally placed to serve as scaning elements.
  • the light passes from B2 in a scanning state and enters the objective lens (OBJ) via a coupling lens l, which functions to bring the back aperture of the objective into a plane conjugate with that of B2.
  • OBJ objective lens
  • the signal from the object being illuminated then traverses this path in reverse and is scanned by the mirror B2. It then passes through the beam splitter BS to the masking slit C where it is brought into focus and traverses the slit. Interfering signals emanating from regions away from the illuminated line are rejected by the slit.
  • the slit is adjustable in width to allow stringent confocal conditions to be imposed (slit narrow) or a brighter image of a less confocal nature to be formed (slit wide).
  • the oscillating mirror Bl in the second Offner unit acts to descan the signal, recreating a stationary two-dimensional image at the mid-plane (MP) which can be viewed directly with the eyepiece or be recorded in a camera.
  • the movement of the convex mirrors Bl and B2 may take any form that scans the whole field, but sweeps of uniform angular velocity in alternate directions are preferable as this reduces dead time and gives an image of uniform brightness.
  • the mirror B2 works in synchronism but antiphase to the oscillating mirror Bl .
  • the beam splitter BS could be either a simple device producing any desired ratio of reflected and transmitted intensities, or alternatively a dichromatic reflector (dichroic) as is conventional in fluorescence microscopy.

Landscapes

  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Ophthalmology & Optometry (AREA)
  • Radiology & Medical Imaging (AREA)
  • Surgery (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

Système de formation d'images à foyer commun s'utilisant en association avec un microscope optique, dans lequel un faisceau lumineux sous forme de fente ou de barre explore l'échantillon, est occulté au moyen d'un masque fixe et remis en action à des fins de visualisation ou d'enregistrement, le système de mise au point et d'exploration étant constitué par des systèmes optiques entièrement réfléchissants (A1, B1 et A2, B2).
EP91913800A 1990-07-28 1991-07-26 Systeme de formation d'images a foyer commun utilise en microscopie Expired - Lifetime EP0541625B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GB9016632 1990-07-28
GB909016632A GB9016632D0 (en) 1990-07-28 1990-07-28 Confocal imaging system for microscopy
PCT/GB1991/001268 WO1992002838A1 (fr) 1990-07-28 1991-07-26 Systeme de formation d'images a foyer commun utilise en microscopie

Publications (2)

Publication Number Publication Date
EP0541625A1 true EP0541625A1 (fr) 1993-05-19
EP0541625B1 EP0541625B1 (fr) 1996-03-06

Family

ID=10679843

Family Applications (1)

Application Number Title Priority Date Filing Date
EP91913800A Expired - Lifetime EP0541625B1 (fr) 1990-07-28 1991-07-26 Systeme de formation d'images a foyer commun utilise en microscopie

Country Status (8)

Country Link
US (2) US5452125A (fr)
EP (1) EP0541625B1 (fr)
JP (1) JPH05509178A (fr)
AT (1) ATE135116T1 (fr)
CA (1) CA2088267A1 (fr)
DE (1) DE69117761T2 (fr)
GB (1) GB9016632D0 (fr)
WO (1) WO1992002838A1 (fr)

Families Citing this family (21)

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Publication number Priority date Publication date Assignee Title
US5923466A (en) * 1993-10-20 1999-07-13 Biophysica Technologies, Inc. Light modulated confocal optical instruments and method
GB9408688D0 (en) * 1994-04-30 1994-06-22 Medical Res Council Scanning confocal optical microscope
US5880465A (en) * 1996-05-31 1999-03-09 Kovex Corporation Scanning confocal microscope with oscillating objective lens
DE19654208C2 (de) * 1996-12-24 2001-05-10 Leica Microsystems Mikroskop
US20030036855A1 (en) * 1998-03-16 2003-02-20 Praelux Incorporated, A Corporation Of New Jersey Method and apparatus for screening chemical compounds
US6388788B1 (en) 1998-03-16 2002-05-14 Praelux, Inc. Method and apparatus for screening chemical compounds
US6548796B1 (en) * 1999-06-23 2003-04-15 Regents Of The University Of Minnesota Confocal macroscope
DE60037682T2 (de) * 2000-09-04 2009-01-02 Hamamatsu Photonics K.K., Hamamatsu Abbildungsvorrichtung
US6309078B1 (en) 2000-12-08 2001-10-30 Axon Instruments, Inc. Wavelength-selective mirror selector
US6856457B2 (en) * 2001-03-27 2005-02-15 Prairie Technologies, Inc. Single and multi-aperture, translationally-coupled confocal microscope
AU2002332802A1 (en) * 2001-08-29 2003-03-10 Musc Foundation For Research Development Line scanning confocal microscope
WO2005083352A1 (fr) * 2004-02-11 2005-09-09 Filmetrics, Inc. Procede et appareil de cartographie d'epaisseur a grande vitesse pour couches minces configurees
US20050237530A1 (en) * 2004-04-26 2005-10-27 Schnittker Mark V Imaging apparatus for small spot optical characterization
US7221449B2 (en) * 2004-06-28 2007-05-22 Applera Corporation Apparatus for assaying fluorophores in a biological sample
DE102004034970A1 (de) * 2004-07-16 2006-02-02 Carl Zeiss Jena Gmbh Lichtrastermikroskop und Verwendung
US7315352B2 (en) 2004-09-02 2008-01-01 Avago Technologies General Ip (Singapore) Pte. Ltd. Offner imaging system with reduced-diameter reflectors
US7173686B2 (en) 2004-09-02 2007-02-06 Agilent Technologies, Inc. Offner imaging system with reduced-diameter reflectors
US7545446B2 (en) * 2005-08-27 2009-06-09 Hewlett-Packard Development Company, L.P. Offner relay for projection system
FR2903785B1 (fr) * 2006-07-13 2008-10-31 Beamind Soc Par Actions Simpli Procede et dispositif de deflexion d'un faisceau lumineux pour balayer une surface cible
KR20140093818A (ko) * 2013-01-17 2014-07-29 삼성전자주식회사 프로파일 측정 시스템
DE102019115931A1 (de) * 2019-06-12 2020-12-17 Carl Zeiss Microscopy Gmbh Optische Anordnung für ein Mikroskop

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Publication number Priority date Publication date Assignee Title
US3669522A (en) * 1970-03-20 1972-06-13 Singer Co Reflective relay optical system for two-axis deflection
US3951546A (en) * 1974-09-26 1976-04-20 The Perkin-Elmer Corporation Three-fold mirror assembly for a scanning projection system
US4170398A (en) * 1978-05-03 1979-10-09 Koester Charles J Scanning microscopic apparatus with three synchronously rotating reflecting surfaces
US5032720A (en) * 1988-04-21 1991-07-16 White John G Confocal imaging system
JP2594470B2 (ja) * 1988-07-06 1997-03-26 メディカル リサーチ カウンシル 無彩色走査装置
US5020891A (en) * 1988-09-14 1991-06-04 Washington University Single aperture confocal scanning biomicroscope and kit for converting single lamp biomicroscope thereto
US5035476A (en) * 1990-06-15 1991-07-30 Hamamatsu Photonics K.K. Confocal laser scanning transmission microscope
US5225923A (en) * 1992-07-09 1993-07-06 General Scanning, Inc. Scanning microscope employing improved scanning mechanism

Non-Patent Citations (1)

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Title
See references of WO9202838A1 *

Also Published As

Publication number Publication date
ATE135116T1 (de) 1996-03-15
DE69117761T2 (de) 1996-09-19
EP0541625B1 (fr) 1996-03-06
CA2088267A1 (fr) 1992-01-29
GB9016632D0 (en) 1990-09-12
US5561554A (en) 1996-10-01
WO1992002838A1 (fr) 1992-02-20
DE69117761D1 (de) 1996-04-11
JPH05509178A (ja) 1993-12-16
US5452125A (en) 1995-09-19

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